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NMR Core Facility Training Course High-Field Biomacromolecular Solution NMR Core Facility National Research Program for Genomic Medicine Topics: Protein Solution NMR Spectroscopy. Instructor: Tai-huang Huang, (Chi-Fon Chang and Winston Wu) Time: Every Saturday 10:00 AM - 1:00 PM for ~ 15 sessions (First session: Oct 2, 2004) Place: B1A room, IBMS, Academia Sinica Pre-requisite: Some prior experience with protein NMR spectroscopy. Textbooks: 1. Lecture by James Keeler on “Understanding NMR spectroscopy” (http://www-keeler.ch.cam.ac.uk/lectures/) 2. Cavanagh, Fairbrother, Palmer, and Skelton: “Protein NMR spectroscopy – Principles and practice” Academic press, 1996. 3. Selected review articles. Curse Content This will be a comprehensive lecture course, focusing on modern high field NMR spectroscopy in solution, with applications to protein structure, dynamics and functional studies. Topics to be covered include: 1. Basic NMR theory, including quantum mechanical and vectorial descriptions of NMR spectroscopy. 2. Basic experimental aspects of NMR: NMR data acquisition and processing. 3. Product operator formalism analysis of pulse programs. 3. Spin dynamics: Coherent selection, phase cycling, gradient enhanced spectroscopy. 4. Heteronuclear multidimensional NMR spectroscopy. 5. Relaxation and protein dynamics. 6. Special topics: TROSY, RDC and reduced dimensionality etc. 7. Applications in protein NMR in solution. Course Outline Lect # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Date 10/2 10/9 10/16 10/23 11/20 11/27 12/4 12/11 12/18 12/25 1/8/05 2/19/05 2/26/05 3/5/05 3/12/05 Topics NMR and Energy level Vector Model Fourier Transform and Data processing How the spectrometer works Product Operator Two dimensional NMR Relaxation Coherence selection and phase cycling NMR Historic Review NMR Historic Review 1924 Pauli proposed the presence of nuclear magnetic moment to explain the hyperfine structure in atomic spectral lines. 1930 Nuclear magnetic moment was detected using refined Stern-Gerlach experiment by Estermann. 1939 Rabi et al. First detected unclear magnetic resonance phenomenon by applying r.f. energy to a beam of hydrogen molecules in the Stern-Gerach set up and observed measurable deflection of the beam. 1946 Purcell et al. at Harvard reported nuclear resonance absorption in paraffin wax. Bloch et al. at Stanford found nuclear resonance in liquid water. 1949 Chemical shift phenomenon was observed. 1952 Nobel prize in Physics was awarded to Purcell and Bloch. 1966 Ernst and Anderson first introduce the Fourier Transform technique into NMR. Late in the 1960s: Solid State NMR was revived due to the effort of Waugh. and associates at MIT. 1966 Ernst and Anderson first introduce the Fourier Transform technique into NMR. Late in the 1960s: 1970 Solid State NMR was revived due to the effort of Waugh. and associates at MIT. Biological application become possible due to the introduction superconducting magnets. NMR imaging was demonstrated. 2D NMR was introduced. 1980s Macromolecular structure determination in solution by NMR was achieved. 1991 Nobel prize in Chemistry was awarded to Richard Ernst. 1990s Continuing development of heteronuclear multi-dimensional NMR permit the determination of protein structure up to 50 KDa. MRI become a major radiological tool in medical diagnostic. 2002 Nobel prize in Chemistry was awarded to Kurt Wuthrich NMR Applications NMR(One is aofversatile and it has applications wide varieties themost, iftool not the most, important analytical in spectroscopic tool.) of subjects in addition to its chemical and biomedical applications, 1. Biomedical applications: including material and quantum computing. a. b. Metabolic studies of biological systems. MRI – diagnostic imaging, flow imaging, chemical shift imaging, functional Isador I. Rabi 1944, Physics Richard R. Ernst 1992, Chemistry Edward M. Purcell 1952, Physics Kurt Wuthrich 2002, Chemistry Paul Lauterbur 2003, Medicine Felix Bloch 1952, Physics Peter Mansfield 2003, Medicine CW NMR 40MHz (1960) 800 MHz Basic Nuclear Spin Interactions 6 Electrons 3 Ho 1 3 Nuclear Spin j 2 Nuclear Spin i 5 4 1 Ho 4 Phonons 4 Dominant interactions: . H = HZ + HD + HS + HQ HZ = Zeeman Interaction HS = Chemical Shielding Interaction. HD = Dipolar Interactions HQ = Quadrupolar Interaction